title: Restrictive Streptomycin Resistance Mutations Decrease the Formation of Attaching and Effacing Lesions in Escherichia coli O157:H7 Strains.
abstract: Streptomycin binds to the bacterial ribosome and disrupts protein synthesis by promoting misreading of mRNA. Restrictive mutations on the ribosomal subunit protein S12 confer a streptomycin resistance (Strr) phenotype and concomitantly increase the accuracy of the decoding process and decrease the rate of translation. Spontaneous Strrmutants ofEscherichia coliO157:H7 have been generated forin vivostudies to promote colonization and to provide a selective marker for this pathogen. The locus of enterocyte effacement (LEE) ofE. coliO157:H7 encodes a type III secretion system (T3SS), which is required for attaching and effacing to the intestinal epithelium. In this study, we observed decreases in both the expression and secretion levels of the T3SS translocated proteins EspA and EspB inE. coliO157:H7 Strrrestrictive mutants, which have K42T or K42I mutations in S12. However, mildly restrictive (K87R) and nonrestrictive (K42R) mutants showed slight or indistinguishable changes in EspA and EspB secretion. Adherence and actin staining assays indicated that restrictive mutations compromised the formation of attaching and effacing lesions inE. coliO157:H7. Therefore, we suggest thatE. coliO157:H7 strains selected for Strrshould be thoroughly characterized beforein vivoandin vitroexperiments that assay for LEE-directed phenotypes and that strains carrying nonrestrictive mutations such as K42R make better surrogates of wild-type strains than those carrying restrictive mutations.
authors.affiliation: Department of Food Science, the Penn State University, University Park, Pennsylvania, USA.
authors.forename: Chun
authors.initials: C
authors.lastname: Chen
authors.affiliation: Departments of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas, USA.
authors.forename: Carla A
authors.initials: CA
authors.lastname: Blumentritt
authors.affiliation: Departments of Microbiology and Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Meredith M
authors.initials: MM
authors.lastname: Curtis
authors.affiliation: Departments of Microbiology and Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Sperandio
authors.affiliation: Departments of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas, USA.|Department of Pathology and Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, Texas, USA.
authors.forename: Alfredo G
authors.initials: AG
authors.lastname: Torres
authors.affiliation: Department of Food Science, the Penn State University, University Park, Pennsylvania, USA egd100@psu.edu.
authors.forename: Edward G
authors.initials: EG
authors.lastname: Dudley